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Scrapie is a fatal, neurodegenerative disease occurring in sheep, goats and mufflons. It is commonly believed that its infectious agent is a protease-resistant form of host-encoded prion protein (PrP). Polymorphism of the coding region of PrP genes has been analyzed in many countries. Polymorphism of codons 136, 154 and 171 has been associated with outbreaks of scrapie. Valine (V) in codon 136, arginine (R) in codon 154, glutamine (Q) and histidine (H) in codon 171 are associated with susceptibility to scrapie while histidine (H) in codon 154 and arginine (R) in codon 171 are associated with resistance to the illness. Alanine (A) in codon 136 is associated with low susceptibility to scrapie.
The study was conducted on six Polish sheep breeds (Polish merino, Zelaznenska sheep, Pomeranian, Kamieniecka sheep, Polish mountain sheep and Polish heath sheep). Individual blood samples were collected and DNA was extracted. The frequencies of alleles (ARR, ARH, ARQ, AHQ, VRQ) and its genotypes of PRNP gene were determined according to the RFLP-PCR method. Depending on the breed and sex, a high diversity of polymorphous forms of the PRNP gene was observed. Higher PRNP gene polymorphism was observed in ewes than in rams. The lowest frequency of alleles was determined in three breeds: Polish merino (ARR, ARQ and VRQ), Polish mountain sheep and Polish heath sheep (ARR, ARQ and AHQ). All five of the determined alleles were observed in Pomeranian and Kamieniecka sheep. The most advantageous frequencies of ARR allele occurred in Polish merino, Kamieniecka and Zelaznenska sheep (particularly in ewes). In Polish mountain sheep and Polish heath sheep the VRQ allele was not observed. In the final results, 11 combinations of genotypes occurred in six Polish sheep breeds. The obtained results indicate a need for the construction of an appropriate breeding program for each breed separately. These breeding programs should enable obtaining higher frequencies of ARR allele occurrence and restricting the incidence of other alleles of the PRNP gene, particularly VRQ allele.
The aims of the study were to detect the polymorphisms of three microsatellite sites: S11, S15, and S24 in the ovine PRNP and to asses their relationship with prion protein (PrP) genotypes in three sheep breeds. To identify 15 PrP genotypes based on polymorphisms at codons 136, 154, and 171, PCR-RFLP analyses were applied. The microsatellite sites were amplified. For each microsatellite two or three alleles per site in homo- and heterozygotic combinations were observed. In the group of scrapie resistant sheep (NSP1), the genotype 152/152 bp of the S11 microsatellite, 179/179 bp of S15, and 144/144 bp of S24 occurred with the highest frequencies. The results of Fisher's exact test showed that associations between PrP genotypes and microsatellite alleles and genotypes in the PRNP were statistically significant (P<0,001). In conclusion, the investigated microsatellite sites may be used as additional genetic markers in prion protein gene variability studies in sheep.
PCR-RFLP method was used to identify polymorphisms in the 136 (A/V), 154 (R/H), and 171 (R/H/Q) codons of the prion protein gene (PRNP) in 174 ewes and rams of the prolific Olkuska sheep breed from three nucleus flocks. Alanine (A) at codon 136, arginine (R) at codon 154, and arginine (R) or glutaminę (Q) at codon 171, responsible for the presence of two alleles (ARR and ARQ) and three genotypes, (ARR/ARR, ARR/ARQ and ARQ/ARQ) were found in the analysed population. In all flocks, ARR allele (the most resistant to scrapie) was the dominant haplotype regardless of sheep sex, and ARR/ARQ genotype had the highest frequency (60.92%). The proportion of the undesirable ARQ/ARQ genotype was only 4.02%. Simulation of genotype distribution for the next generations showed that the mating of ewes with ARR/ARR genotype rams will cause this genotype to appear in 99% of Olkuska ewes already in the sixth generation. However, the study showed no relationship between genotype in the PRPN locus and prolificacy potential of the ewes.
The activity of the National Reference Laboratory for a given disease encompasses not only diagnostics but also research and instruction. In the case of transmissible spongiform encephalopathies this range of activities constantly expands along with the accumulation of the data on prion protein characteristics, pathogenesis of spongiform encephalopathies and the threat to consumer health. Besides the standard task, which is confirmatory testing, the reference laboratory should organize instruction, verify diagnostic methods, control the quality of diagnostic reagents, cooperate with the community reference laboratories, participate and arrange comparative tests, as well as take part in the investigations of expert working groups. All the above mentioned aspects of the activity of the National reference laboratory for BSE and scrapie are presented in this review.
Isolation of atypical pestivirus from clinically affected calves in Italy. Experimental studies of foot-and-mouth disease transmission in cattle and the implications for control, controversial preemptive control measures may be unnecessary. Different gene expression profiles of bovine brains infected with classical and atypical BSE indicate possible different pathogenesis or origin of the disease. Emergence of classical BSE from atypical BSE-H supports the hypothesis that this type may be at the origin of the food-borne BSE epizooty. First cases of atypical scrapie in Poland. Two drugs that prevent the differentiation of Tʜ17 cells which mediate autoimmune disorders. Local proliferation of macrophages in Tʜ2 inflammation. Macrophages can enhance tumour resistance to chemotherapy.
The objective of this research was to determine allele and genotype frequencies of the prion protein gene (PRNP) in a flock of sheep and categorise thereof into groups by susceptibility to scrapie. Additionally, a mating simulation was conducted, which aimed at decreasing sheep susceptibility to scrapie. Sheep of Kamieniecka breed from a single flock were investigated. Polymorphism in the PrP gene was identified with the use of the PCR-RFLP method. Four of the five alleles were found - ARR, ARQ, ARU. and VRQ, in nine different combinations - ARR/ARR, ARR/ARQ, ARQ/ARQ, ARR/ARH, ARH/VRQ, ARR/VRQ, ARQ/ARH, ARH/ARH, and ARQ/VRQ. The most frequent allele was the one being the most desired, namely ARR, (63.2%), and the least frequent was VRQ (5.4%). Mating simulation aiming at increasing the frequency of the ARR allele in the examined sheep group demonstrated that in the F₆ generation it is possible to obtain as many as 97.7% of homozygotic animals having the lowest susceptibility to scrapie.
The use of BSE-contaminated meat and bone meal for feeding sheep and goats may be the cause of transmitting the BSE agent into small ruminant populations. Experimental studies have shown that sheep which are fed cattle brain homogenates from BSE cases can succumb to a BSE-like disease. The distribution of BSE agents in these sheep is similar to scrapie and a wider range of organs is affected when compared with BSE in cattle. Thus, the possibility of crossing the species barrier between sheep and man cannot be excluded (as has been shown with BSE and its variant - Creutzfeldt-Jakob disease in man). The paper describes implementing one of the approved immuno-blot methods used for discriminating between BSE and scrapie in a small ruminant population. The use of two monoclonal antibodies of which only one reacts with scrapie and atypical scrapie samples facilitates differentiation between BSE and scrapie positive samples from sheep and goats.
The review describes current diagnostic techniques used for TSEs diagnosis with emphasis on BSE and scrapie in sheep. The clinical signs and their reliability are presented along with the review of laboratory tests. Special emphasis has been placed on the diagnostic reliability of clinical signs including their sensitivity and specificity. Histopatological examination for BSE is discussed with guidelines for diagnostic criteria on single section examination. Other techniques for PrP detection like: SAF detection, immunohistochemistry and western-blotting have also been presented. The review describes the development of novel methods for BSE diagnosis in live animals using cerobrospinal fluid and urine.
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